Fig. 2: The amyloid nature within the p53 family phase separation and transition.

Microscopy images were captured, presenting the Congo red (CR) fluorescence and bright-field channels of (a) p53C, (b) M237I, (c) p63C, and (d) p73C at 4 and 37 °C. White arrows indicate the presence of p53C and M237I aggregates. Scale bar: 20 μm; Line plots showing the area under the light scattering curve as a function of increasing temperatures (4, 25, and 37 °C) of (e) p53C (green), (f) M237I (gray), (g) p63C (light red), and (h) p73C (light blue) in the absence (colored circles) and the presence (colored squares) of PEG. Plots in (g, h) show two segments in the y-axis to highlight negligible scattering. Line plots with the filled area showing turbidity at 600 nm as a function of increasing concentrations of (i) p53C (green palette), (j) M237I (gray palette), (k) p63C (red palette), and (l) p73C (blue palette) at 4, 25, and 37 °C. Plots in (k, l) show two segments in the y-axis to highlight negligible turbidity. Red and blue palettes are not depicted in (k, l) due to similar values across studied temperatures. Scatter dot plots showing the thioflavin T fluorescence as a function of increasing temperatures (4, 25, and 37 °C) of (m) p53C, (n) M237I, (o) p63C, and (p) p73C. The color palette is the same as in (i–l). Plots show two segments in the y-axis to evidence large changes in the thioflavin T signal across studied proteins. The data are shown as the mean ± s.e.m. of n = 3 independent experiments using the same protein batch.